- 标题
- 摘要
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Peptic Fluorescent “Signal-On” and “Signal-Off” Sensors Utilized for the Detection Protein Post-Translational Modifications
摘要： Protein post-translational modifications (PTMs) are typically enzyme-catalyzed events generating functional diversi?cation of proteome; thus, multiple PTM enzymes have been validated as potential drug targets. We have previously introduced energy-transfer-based signal-modulation method called quenching resonance energy transfer (QRET), and utilize it to monitor PTM addition or removal using the developed peptide-break technology. Now we have reinvented the QRET technology, and as a model, we introduced the tunable ?uorescent “signal-on” and “signal-o?” detection scheme in the peptide-break PTM detection. Taking the advantage of time-resolved ?uorescence-based single-label detection technology, we were able to select the signal direction upon PTM addition or removal by simply introducing di?erent soluble Eu3+-signal-modulating molecule. This enables the selection of positive signal change upon measurable event, without any additional labeling steps, changes in assay condition or Eu3+-reporter. The concept functionality was demonstrated with four Eu3+-signal modulators in a high-throughput compatible kinase and phosphatase assays using signal-on and signal-o? readout at 615 nm or time-resolved Fo?rster resonance energy transfer at 665 nm. Our data suggest that the introduced signal modulation methodology provides a transitional ?uorescence-based single-label detection concept not limited only to PTM detection.
关键词： time-resolved fluorescence,signal-off,high-throughput screening,peptide-break technology,protein post-translational modifications,FRET,signal-on
更新于2025-11-19 16:56:35
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Supersandwich Nanowire/Quantum Dots Sensitization Structure-Based Photoelectrochemical a??Signal-Ona?? Platform for Ultrasensitive Detection of Thrombin
摘要： A new photoelectrochemical (PEC) “signal-on” sensing platform based on photoactive material Bi2O3-ZnO and CdS quantum dots (QDs) sensitizer were fabricated for ultrasensitive determination of thrombin by constructing supersandwich nanowires. The CdS/ZnO/Bi2O3 sensitization structure with excellent energy level arrangement remarkably improved photoelectric conversion efficiency due to efficient separation of electron-hole. Moreover, the DNA supersandwich nanowire is ingeniously synthesized in one step by simple dislocation hybridization, which could carry a large amount of sensitized material CdS QDs. More importantly, with Exo III-assisted multiple amplification, the proposed “signal-on” platform demonstrated a detection range of 10 fM to 1 μM with the detection limit of 1.41 fM for thrombin. Impressively, the PEC platform can successfully detect human serum samples with good accuracy. Above all, the CdS/ZnO/Bi2O3 sensitization photoelectric biosensing platform by using DNA nanowire in combination with Exo III-multiple amplification open new sensitized amplification paths for supersensitive biosensing and bioanalysis.
关键词： photoelectrochemical,CdS quantum dots,signal-on,thrombin detection,Bi2O3-ZnO,supersandwich nanowires,Exo III-assisted multiple amplification
更新于2025-09-19 17:13:59